Effects of human papillomavirus type 16 E5 deletion mutants on epithelial morphology: functional characterization of each transmembrane domain
Effects of human papillomavirus type 16 E5 deletion mutants on epithelial morphology: functional characterization of each transmembrane domain
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DOI：
10.1099/vir.0.016295-0
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发表时间：
2010-02-01
影响因子：
3.8
通讯作者：
Campo, M. Saveria
中科院分区：
文献类型：
--
作者：
Barbaresi, Silvia;Cortese, Marc S.;Campo, M. Saveria
人乳头瘤病毒16型（HPV-16）是宫颈癌的病因。HPV基因组编码三种转化蛋白，E5、E6和E7，E6和E7是HPV的主要转化蛋白，而E5的作用仍然知之甚少。使用三维器官型筏培养，我们表明，HaCaT人角质形成细胞表达HPV-16 E5形成一个非常扰动的上皮，同时过度角化的一些细胞和其他细胞的分化缺陷。基底层受到干扰，许多细胞侵入胶原基质。分化层中的许多细胞显示出基底细胞的特征：通过细胞周期的进展、细胞角蛋白14的表达、细胞角蛋白1的缺乏和基质金属蛋白酶（MMP）的产生。使用包含E5的三个疏水结构域的缺失突变体，我们将促进基质侵袭的能力分配给第一个疏水结构域，并将诱导MMP 9的能力分配给C末端的四个氨基酸。我们还表明，侵袭和MMP 9的产生可以分离，因为仍然能够侵袭的突变体不产生MMP 9，反之亦然。
Human papillomavirus type 16 (HPV-16) is the cause of cervical cancer. The HPV genome encodes three transforming proteins, E5, E6 and E7, E6 and E7 are the main transforming proteins of HPV, while the role of E5 is still poorly understood. Using three dimensional organotypic raft cultures we show that HaCaT human keratinocytes expressing HPV-16 E5 form a very perturbed epithelium, with simultaneous hyperkeratinization of some cells and defective differentiation of other cells. The basal layer is disturbed and many cells invade the collagen matrix. Many cells among the differentiated layers show characteristics of basal cells: progression through the cell cycle, expression of cytokeratin 14, lack of cytokeratin 1 and production of matrix metalloproteases (MMP). Using deletion mutants which encompass the three hydrophobic domains of E5, we have assigned the ability to promote invasion of the matrix to the first hydrophobic domain, and the capacity to induce MMP9 to the C-terminal four amino acids. We also show that invasion and production of MMP9 can be dissociated, as mutants that are still capable of invasion do not produce MMP9 and vice versa.